Dataset Information

Proteomic analysis of Caenorhabditis elegans wound model reveals novel molecular players involved in repair

ABSTRACT: Now a days, wound healing is becoming a global threat that impact economy of the country severely. Though the steps involved in wound healing are well characterised, direct therapeutics for the accelerated wound healing is comparatively less. This is solely because of the incomplete mechanism of healing and the lack of knowledge on molecular players involved in wound healing. Hence, in the present study, we have investigated the molecular players involved in wound healing process using a versatile model organism Caenorhabditis elegans through proteomic analyses. Especially, we have employed the high through put proteomic analyses tools such as 2-D GE and LCMS/MS analysis to uncover the molecular players involved in wound healing. As a result of LC-MS/MS analysis at 0 and 24 h, a total of 435 proteins were regulated in both unwounded and wounded conditions in which 13 and 10 proteins were significantly differential regulated. Bioinformatics analyses result suggested that molecular players involved in cortical actin cytoskeleton organization, protein phosphatase binding, and proton transporting ATP synthase activity were identified at 0 h; skeletal muscle myosin thick filament assembly, actin filament depolymerization were identified at 24 h.

INSTRUMENT(S): 6545 Q-TOF LC/MS

ORGANISM(S): Caenorhabditis Elegans

TISSUE(S): Whole Body

DISEASE(S): Wounds And Injuries

SUBMITTER: Krishnaswamy Balamurugan

LAB HEAD: Dr. KB Lab (Host Pathogen Interaction Lab)

PROVIDER: PXD024744 | Pride | 2021-04-16

REPOSITORIES: Pride

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Proteomic analysis of Caenorhabditis elegans wound model reveals novel molecular players involved in repair.

Pooranachithra Murugesan M Satheesh Kumar Chelladurai C Bhaskar James Prabhanand JP Venkateswaran Krishnan K Ravichandiran Velayutham V Balamurugan Krishnaswamy K

Journal of proteomics 20210405

Wound repair is a multistep process which involves coordination of multiple molecular players from different cell types and pathways. Though the cellular processes that are taking place in order to repair damage is already known, molecular players involved in crucial pathways are still scarce. In this regard, the present study intends to uncover crucial players that are involved in the central repair events through proteomics approach which included 2-D GE and LC-MS/MS using Caenorhabditis elega ...[more]

PMID: 33831597

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Project description:Wound healing is an essential homeostatic mechanism that maintains the epithelial barrier integrity after tissue damage. Although we know the main events participating in the healing of a wound, many of the underlying molecular mechanisms remain unclear. Genetically amenable systems, such as wound healing in Drosophila imaginal discs, do not model all aspects of the repair process, but allow exploring many unanswered features of the healing response; e.g., which are the signal(s) responsible for initiating tissue remodeling? How is the sealing of the epithelia achieved? Or which are the inhibitory cues that cancel the healing machinery upon completion? Answering these and other questions demands in first place the identification and functional analysis of wound-specific genes. A variety of different microarray analyses of murine and humans have identified characteristic profiles of gene expression at the wound site, however, very few functional studies in healing regulation have been carried out. We developed an experimentally controlled method to culture imaginal discs that allows live imaging and biochemical analysis and is healing-permissive. Employing this approach, we performed a comparative genome-wide profiling between those Drosophila imaginal cells actively involved in healing versus their non-engaged siblings. This lets us identify a set of potential wound-specific genes. Importantly, besides identifying and categorizing new genes, we functionally tested many of their gene products by genetic interference and overexpression in a healing assay. This non-saturated analysis defines a relevant set of new genes whose changes in expression levels are functionally significant for proper tissue repair. There is promise that our newly identified wound-healing genes will guide future work in the more complex mammalian wound response. We developed a healing-permissive in vitro culture system for fly imaginal discs: we used one-channel microarrays for comparing healing-engaged cells (showing activation of the JNK signaling cascade) to cells not participating in healing (silent JNK activity) in wounded wing imaginal discs in culture. Employing this method, we aimed detecting the relevant genes involved in disc healing through microarray analysis. We compared cells actively involved in healing to those not involved and identified a whole set of upregulated or downregulated genes. They were annotated, clustered by expression profiles, chromosomal locations and presumptive functions. Most importantly, we functionally tested them genetically in a healing assay.

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